Effect of fluoroquinolone exposure on the proteome of Salmonella enterica serovar Typhimurium
Effect of fluoroquinolone exposure on the proteome of Salmonella enterica serovar Typhimurium
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DOI：
10.1093/jac/dkl413
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发表时间：
2006-12-01
影响因子：
5.2
通讯作者：
Woodward, Martin J.
中科院分区：
文献类型：
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作者：
Coldham, Nick G.;Randall, Luke P.;Woodward, Martin J.
目的：用蛋白质组学方法研究了沙门氏菌对氟喹诺酮类抗生素的生理反应。方法：用环丙沙星（0.03和0.008 mg/L; 2x和0.5x MIC）和恩诺沙星（0.03 mg/L）处理肉汤培养物后，从菌株SL 1344和多重抗生素耐药（MAR）突变株制备蛋白质组。通过二维HPLC-MSn以及暴露于环丙沙星后通过二维凝胶电泳（2D-GE）测定蛋白质表达。蛋白质的数目通过来自野生型菌株的对照培养物的2D-GE检测到的（平均值+/- SD）显著高于对照培养物。用环丙沙星（0.03 mg/L）治疗后，从296 +/- 77降至153 +/- 36（P < 0.05）。凸起表达式17种蛋白质的表达也有显著性差异（P < 0.05），（P < 0.0001）。二维HPLC-MSn分析提供了更高的蛋白质组覆盖率，检测到787 +/- 50个蛋白质，环丙沙星（0.03mg/L）使其降低至560 ± 14（P < 0.005）。观察到43种蛋白质的表达增加，其中包括通过2D-GE检测到的蛋白质以及外排泵蛋白AcrB。与未处理的野生型相比，MAR突变体中AcrAB/TolC外排泵的基础表达升高，并且在用环丙沙星（0.03 mg/L）处理后增强。F1 F0-ATP合成酶和Imp仅在突变体中升高，当用环丙沙星处理时。结论：这些研究表明，AcrAB/TolC的表达增加与抗性相关，而其他增加，如F1 F0-ATP合成酶和Imp，是对氟喹诺酮的反应。
Objectives: The physiological response of Salmonella enterica serovar Typhimurium to fluoroquinolone antibiotics was investigated using proteomic methods.Methods: Proteomes were prepared from strain SL1344 following treatment of broth cultures with ciprofloxacin (0.03 and 0.008 mg/L; 2x and 0.5x MIC) and enrofloxacin (0.03 mg/L) and from a multiple antibiotic resistant (MAR) mutant. Protein expression was determined by two-dimensional HPLC-MSn and also after exposure to ciprofloxacin by two-dimensional gel electrophoresis (2D-GE).Results: The number of proteins (mean +/- SD) detected by 2D-GE derived from control cultures of the wild-type strain was significantly (P < 0.05) reduced from 296 +/- 77 to 153 +/- 36 following treatment with ciprofloxacin (0.03 mg/L). Raised expression (P < 0.05) of 17 proteins was also detected, and increases of up to 8-fold (P < 0.0001) were observed for subunits of F1F0-ATP synthase, TolC and Imp. Analysis by two-dimensional HPLC-MSn provided higher proteome coverage with 787 +/- 50 proteins detected, which was reduced (P < 0.005) to 560 +/- 14 by ciprofloxacin (0.03 mg/L). Increased expression of 43 proteins was observed which included those detected by 2D-GE and additionally the efflux pump protein AcrB. The basal expression of the AcrAB/TolC efflux pump was elevated in the MAR mutant compared with the untreated wild-type and augmented following treatment with ciprofloxacin (0.03 mg/L). F1F0-ATP synthase and Imp were only elevated in the mutant when treated with ciprofloxacin.Conclusions: These studies suggest that increased expression of AcrAB/TolC was associated with resistance while other increases, such as in F1F0-ATP synthase and Imp, were a response to fluoroquinolone.